Abstract

Despite the enormous replication potential of the human liver, there are currently no culture
systems available that sustain hepatocyte replication and/or function in vitro. We have shown
previously that single mouse Lgr5+ liver stem cells can be expanded as epithelial organoids in
vitro and can be differentiated into functional hepatocytes in vitro and in vivo. We now
describe conditions allowing long-term expansion of adult bile duct-derived bi-potent
progenitor cells from human liver. The expanded cells are highly stable at the chromosome
and structural level, while single base changes occur at very low rates. The cells can readily
be converted into functional hepatocytes in vitro and upon transplantation in vivo. Organoids
from α1-antitrypsin deficiency and Alagille Syndrome patients mirror the in vivo pathology.
Clonal long-term expansion of primary adult liver stem cells opens up experimental avenues
for disease modeling, toxicology studies, regenerative medicine and gene therapy.

Sponsorship

This work was supported by grants to MH (EU/236954) and to HC (The United European Gastroenterology Federation
(UEGF) Research Prize 2010, EU/232814-StemCellMark and NWO/116002008). MH is
supported by The Wellcome Trust Sir Henry Dale fellowship. The Rspo cell line was kindly
provided by Dr. Calvin Kuo.